CONTROL OF TRANSIENT EXPRESSION OF CHIMERIC GENES BY GIBBERELLIC-ACID AND ABSCISIC-ACID IN PROTOPLASTS PREPARED FROM MATURE BARLEY ALEURONE LAYERS
CONTROL OF TRANSIENT EXPRESSION OF CHIMERIC GENES BY GIBBERELLIC-ACID AND ABSCISIC-ACID IN PROTOPLASTS PREPARED FROM MATURE BARLEY ALEURONE LAYERS
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DOI：
10.1007/bf00023435
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发表时间：
1991-04-01
影响因子：
5.1
通讯作者：
CLOSE, TJ
中科院分区：
文献类型：
--
作者：
JACOBSEN, JV;CLOSE, TJ
赤霉素（GA_3）和脱落酸（阿坝）控制大麦糊粉层细胞α-淀粉酶基因的转录。 这种控制很可能是通过基因启动子区的顺式作用酶反应元件来实现的。 为了进一步确定这些元素，我们已经开发了程序，获得从成熟大麦糊粉层制备的原生质体中的嵌合基因的瞬时表达。 这些细胞特异性表达具有异源组成型启动子和具有异源和同源GA 3-和ABA-调节启动子的构建体。 该系统为基因调控研究提供了很大的潜力，特别是对同源基因的功能研究。利用该系统对大麦α-淀粉酶基因进行了功能分析。 将来自高pI α-淀粉酶基因的2050 bp片段与报告基因（GUS）融合，并检查其表达的控制。 该启动子片段的缺失分析表明，主要的GA-和ABA-响应元件发生在转录起始位点上游174和41 bp之间。
Gibberellic acid (GA3) and abscisic acid (ABA) control the transcription of alpha-amylase genes in barley aleurone cells. This control is likely to be exerted through cis-acting hormone-responsive elements in the promoter region of the gene. In order to further define these elements, we have developed procedures for obtaining transient expression of chimaeric genes in protoplasts prepared from mature barley aleurone layers. Constructs with heterologous constitutive promoters and with heterologous and homologous GA3- and ABA-regulated promoters were expressed specifically by these cells. This system would appear to offer great potential in gene regulation studies especially for hormonally regulated homologous genes.Functional analysis of a barley alpha-amylase gene has been performed using this system. A 2050 bp fragment from a high-pI alpha-amylase gene was fused to a reporter gene (GUS) and control of its expression was examined. Deletion analysis of this promoter fragment showed that major GA- and ABA-responsive elements occurred between 174 and 41 bp upstream from the transcription initiation site.